Target (protein/gene name): NCBI #: Protein ID (NP or XP #) or Wolbachia#: Organism (including strain): Etiologic Risk Group (see link below):
Disease Information (sort of like the Intro to your Mini Research Write up): Link to TDR Targets page (if present): Link to Gene Database page NCBI:
Essentiality of this protein: Is it a monomer or multimer as biological unit:
Complex of proteins?:
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): *EC#: Link to BRENDA EC# page: Enzyme Assay information: A continuous, spectrophotometric assay is available. Link to paper. -- links to assay reagents (substrates) pages. --- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model) PDB #: 2VPQ Similarity to Human: In eukaryotes, the BC, CT and BCCD subunits reside in a single polypeptide chain. In bacteria, ACC assembles as a multisubunit complex with each of the reactions performed by a separate enzyme. These differences in tertiary structure can provide for selective inhibitors.
Current Inhibitors: One weak inhibitor has been discovered. Expression Information (has it been expressed in bacterial cells): It can and has been expressed in E. coli. Expressed in cytoplasm. ACC is essential for growth, is expressed at all times, expression is also very regulated to maintain the correct stoichiometry of subunits. Purification Method: Standard nickel affinity column chromatography can be used. Image of protein (PyMol with features delineated and shown separately):
2VPQ_bio_r_500.jpg
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): Chain A Sequence: MKKVLIANRGEIAVRIIRACRDLGIQTVAIYSEGDKDALHTQIADEAYCVGPTLSKDSYLNIPNILSIATSTGCDGVHPGYGFLAENADFAELCEACQLKFIGPSYQSIQKMGIKDVAKAEMIKANVPVVPGSDGLMKDVSEAKKIAKKIGYPVIIKATGGGGKGIRVARDEKELETGFRMTEQEA QTAFGNGGLYMEKFIENFRHIEIQIVGDSYGNVIHLGERDCTIQRRMQKLVEEAPSPILDDETRREMGNAAVRAAKAVNYENAGTIEFIYDLNDNKFYFMEMNTRIQVEHPVTEMVTGIDLVKLQLQVAMGDVLPYKQEDIKLTGHAIEFRINAENPYKNFMPSPGKIEQYLAPGGYGVRIESA CYTNYTIPPYYDSMVAKLIIHEPTRDEAIMAGIRALSEFVVLGIDTTIPFHIKLLNNDIFRSGKFNTNFLEQNSIMNDEG Chain B Sequence: MKKVLIANRGEIAVRIIRACRDLGIQTVAIYSEGDKDALHTQIADEAYCVGPTLSKDSYLNIPNILSIATSTGCDGVHPGYGFLAENADFAELCEACQLKFIGPSYQSIQKMGIKDVAKAEMIKANVPVVPGSDGLMKDVSEAKKIAKKIGYPVIIKATAGGGGKGIRVARDEKELETGFRMTEQE AQTAFGNGGLYMEKFIENFRHIEIQIVGDSYGNVIHLGERDCTIQRRMQKLVEEAPSPILDDETRREMGNAAVRAAKAVNYENAGTIEFIYDLNDNKFYFMEMNTRIQVEHPVTEMVTGIDLVKLQLQVAMGDVLPYKQEDIKLTGHAIEFRINAENPYKNFMPSPGKIEQYLAPGGYGVRIES ACYTNYTIPPYYDSMVAKLIIHEPTRDEAIMAGIRALSEFVVLGIDTTIPFHIKLLNNDIFRSGKFNTNFLEQNSIMNDEG Length of your protein in Amino Acids: 451 amino acids. Molecular Weight of your protein in kiloDaltons: 50.05 kDA Molar Extinction coefficient of your protein at 280 nm wavelength: 28685 mol^-1cm-1 TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
TMPRED.20685.9091.gif
*CDS Gene Sequence (paste as text only): ATGAAAAAGGTTTTAATTGCAAACCGCGGTGAAATCGCAGTTAGGATTATTCGCGCTTGTCGTGATTTAGGCATCCAAACTGTTGCAATCTATTCTGAAGGGGATAAAGATGCGCTACATACTCAAATTGCTGATGAAGCATATTGCGTAGGTCCCACTTTGTCTAAAGATTCATATTTAAATATTCCGAACATCTTATCT ATTGCAACTTCTACAGGTTGTGATGGCGTCATCCGGGTTATGGCTTTTTAGCTGAAAATGCTGACTTTGCAGAATTATGCGAAGCATGCCAATTGAAGTTCATTGACCAAGTTATCAATCTATCCAAAAAATGGGTATCAAAGATGTTGCTAAGGCAGAAATGATCAAAGCCAATGTTCCAGTTGTTCCTGGTAGTGAC GGTTTAATGAAAGACGTCTCAGAAGCTAAGAAAATCGCTAAGAAAATTGGCTATCCGGTCATCATTAAAGCTACTGCTGGTGGTGGCGGAAAAGGTATCCGTGTTGCTCGTGATGAAAAAGAACTTGAAACTGGCTTCCGAATGACAGAACAAGAAGCTCAAACTGCATTTGGTAATGGTGGACTTTATATGGAGA AATTCATCGAAAACTTCCGCCATATTGAAATCCAAATTGTTGGGGACAGCTATGGTAATGTAATTCATTTAGGAGAACGTGATTGTACAATTCAAAGACGTATGCAGAAATTAGTGGAAGAAGCACCTTCCCCAATTTTAGATGATGAAACACGTCGTGAAATGGGAAATGCCGCAGTTCGTGCAGCGAAAGCTGTA AATTATGAAAATGCGGGAACAATTGAGTTTATATATGATTTAAATGATAATAAATTTTATTTTATGGAAATGAATACACGTATTCAAGTAGAACATCCTGTAACTGAAATGGTAACAGGAATTGATTTAGTTAAATTACAATTACAAGTTGCTATGGGTGACGTGTTACCGTATAAACAAGAAGATATTAAATAACAGGACA CGCAATTGAATTTAGAATTAATGCTGAAAATCCTTACAAGAACTTTATGCCATCACCAGGTAAAATTGAGCAATATCTTGCACCAGGTGGATATGGTGTTCGAATAGAGTCAGCATGTTATACTAATTATACGATACCGCCATATTATGATTCGATGGTAGCGAAATTAATCATACATGAACCGACACGAGATGAAGCGAT TATGGCTGGCATTCGTGCATTAAGTGAATTTGTGGTTCTTGGTATTGATACAACTATTCCATTCCATATTAAATTATTGAATAACGATATATTTAGAAGCGGTAAATTTAATACAAACTTTTTAGAGCAAAATAGCATTATGAATGATGAAGGTTAA
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
*GC% Content for gene:
output
*GC% Content for gene (codon optimized):
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences): **
NCBI #:
Protein ID (NP or XP #) or Wolbachia#:
Organism (including strain):
Etiologic Risk Group (see link below):
Disease Information (sort of like the Intro to your Mini Research Write up):
Link to TDR Targets page (if present):
Link to Gene Database page NCBI:
Essentiality of this protein:
Is it a monomer or multimer as biological unit:
Complex of proteins?:
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
*EC#:
Link to BRENDA EC# page:
Enzyme Assay information:
A continuous, spectrophotometric assay is available. Link to paper.
-- links to assay reagents (substrates) pages.
--- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model)
PDB #: 2VPQ
Similarity to Human:
In eukaryotes, the BC, CT and BCCD subunits reside in a single polypeptide chain. In bacteria, ACC assembles as a multisubunit complex with each of the reactions performed by a separate enzyme. These differences in tertiary structure can provide for selective inhibitors.
Current Inhibitors:
One weak inhibitor has been discovered.
Expression Information (has it been expressed in bacterial cells): It can and has been expressed in E. coli. Expressed in cytoplasm. ACC is essential for growth, is expressed at all times, expression is also very regulated to maintain the correct stoichiometry of subunits.
Purification Method: Standard nickel affinity column chromatography can be used.
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
Chain A Sequence:
MKKVLIANRGEIAVRIIRACRDLGIQTVAIYSEGDKDALHTQIADEAYCVGPTLSKDSYLNIPNILSIATSTGCDGVHPGYGFLAENADFAELCEACQLKFIGPSYQSIQKMGIKDVAKAEMIKANVPVVPGSDGLMKDVSEAKKIAKKIGYPVIIKATGGGGKGIRVARDEKELETGFRMTEQEA
QTAFGNGGLYMEKFIENFRHIEIQIVGDSYGNVIHLGERDCTIQRRMQKLVEEAPSPILDDETRREMGNAAVRAAKAVNYENAGTIEFIYDLNDNKFYFMEMNTRIQVEHPVTEMVTGIDLVKLQLQVAMGDVLPYKQEDIKLTGHAIEFRINAENPYKNFMPSPGKIEQYLAPGGYGVRIESA
CYTNYTIPPYYDSMVAKLIIHEPTRDEAIMAGIRALSEFVVLGIDTTIPFHIKLLNNDIFRSGKFNTNFLEQNSIMNDEG
Chain B Sequence:
MKKVLIANRGEIAVRIIRACRDLGIQTVAIYSEGDKDALHTQIADEAYCVGPTLSKDSYLNIPNILSIATSTGCDGVHPGYGFLAENADFAELCEACQLKFIGPSYQSIQKMGIKDVAKAEMIKANVPVVPGSDGLMKDVSEAKKIAKKIGYPVIIKATAGGGGKGIRVARDEKELETGFRMTEQE
AQTAFGNGGLYMEKFIENFRHIEIQIVGDSYGNVIHLGERDCTIQRRMQKLVEEAPSPILDDETRREMGNAAVRAAKAVNYENAGTIEFIYDLNDNKFYFMEMNTRIQVEHPVTEMVTGIDLVKLQLQVAMGDVLPYKQEDIKLTGHAIEFRINAENPYKNFMPSPGKIEQYLAPGGYGVRIES
ACYTNYTIPPYYDSMVAKLIIHEPTRDEAIMAGIRALSEFVVLGIDTTIPFHIKLLNNDIFRSGKFNTNFLEQNSIMNDEG
Length of your protein in Amino Acids: 451 amino acids.
Molecular Weight of your protein in kiloDaltons: 50.05 kDA
Molar Extinction coefficient of your protein at 280 nm wavelength: 28685 mol^-1cm-1
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
*CDS Gene Sequence (paste as text only):
ATGAAAAAGGTTTTAATTGCAAACCGCGGTGAAATCGCAGTTAGGATTATTCGCGCTTGTCGTGATTTAGGCATCCAAACTGTTGCAATCTATTCTGAAGGGGATAAAGATGCGCTACATACTCAAATTGCTGATGAAGCATATTGCGTAGGTCCCACTTTGTCTAAAGATTCATATTTAAATATTCCGAACATCTTATCT
ATTGCAACTTCTACAGGTTGTGATGGCGTCATCCGGGTTATGGCTTTTTAGCTGAAAATGCTGACTTTGCAGAATTATGCGAAGCATGCCAATTGAAGTTCATTGACCAAGTTATCAATCTATCCAAAAAATGGGTATCAAAGATGTTGCTAAGGCAGAAATGATCAAAGCCAATGTTCCAGTTGTTCCTGGTAGTGAC
GGTTTAATGAAAGACGTCTCAGAAGCTAAGAAAATCGCTAAGAAAATTGGCTATCCGGTCATCATTAAAGCTACTGCTGGTGGTGGCGGAAAAGGTATCCGTGTTGCTCGTGATGAAAAAGAACTTGAAACTGGCTTCCGAATGACAGAACAAGAAGCTCAAACTGCATTTGGTAATGGTGGACTTTATATGGAGA
AATTCATCGAAAACTTCCGCCATATTGAAATCCAAATTGTTGGGGACAGCTATGGTAATGTAATTCATTTAGGAGAACGTGATTGTACAATTCAAAGACGTATGCAGAAATTAGTGGAAGAAGCACCTTCCCCAATTTTAGATGATGAAACACGTCGTGAAATGGGAAATGCCGCAGTTCGTGCAGCGAAAGCTGTA
AATTATGAAAATGCGGGAACAATTGAGTTTATATATGATTTAAATGATAATAAATTTTATTTTATGGAAATGAATACACGTATTCAAGTAGAACATCCTGTAACTGAAATGGTAACAGGAATTGATTTAGTTAAATTACAATTACAAGTTGCTATGGGTGACGTGTTACCGTATAAACAAGAAGATATTAAATAACAGGACA
CGCAATTGAATTTAGAATTAATGCTGAAAATCCTTACAAGAACTTTATGCCATCACCAGGTAAAATTGAGCAATATCTTGCACCAGGTGGATATGGTGTTCGAATAGAGTCAGCATGTTATACTAATTATACGATACCGCCATATTATGATTCGATGGTAGCGAAATTAATCATACATGAACCGACACGAGATGAAGCGAT
TATGGCTGGCATTCGTGCATTAAGTGAATTTGTGGTTCTTGGTATTGATACAACTATTCCATTCCATATTAAATTATTGAATAACGATATATTTAGAAGCGGTAAATTTAATACAAACTTTTTAGAGCAAAATAGCATTATGAATGATGAAGGTTAA
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
*GC% Content for gene:
output
*GC% Content for gene (codon optimized):
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
**